采用成分明确的体外培养系统产生牛体外受精胚胎,结合荧光免疫定位的方法,利用激光共聚焦成像系统研究了在牛卵母细胞及不同发育阶段早期胚胎中IGF-Ⅰ及其受体的表达。结果表明,IGF-Ⅰ及其受体从未成熟的卵母细胞、成熟的卵母细胞到早期胚胎发育的各个阶段均有表达,在COC的颗粒细胞中也有表达.IGF-Ⅰ在成熟卵母细胞中主要定位于细胞膜附近,在未成熟卵母细胞、2细胞期胚胎中分布于细胞膜及膜下区域,在8细胞期胚胎和桑椹胚中分布于整个细胞质中,在囊胚阶段主要位于滋胚层,内细胞团细胞有少量的表达.IGF-IR在未成熟卵母细胞和成熟卵母细胞以及2细胞、8细胞期胚胎中集中分布于卵裂球远离胚胎中心的细胞膜上,在桑椹胚期时遍布于整个胚胎细胞的细胞质中,在囊胚期时主要分布在滋胚层,内细胞团细胞有少量表达.
The aim of the study was to illuminate the expression of IGF-Ⅰ and it's receptor in bovine oocytes and preimplantation embryos derived from IVF by using indirect immunofluorescence and laser scanning confocal microscopy. The results showed that IGF-I and IGF-IR were detected in COC, immature oocytes, mature oocytes and embryos in 2-cell, 8-cell, morula and blastocyst stages. In IGF-Ⅰ experiments immunofluorescence of mature oocytes was observed on the whole cell membrane. Strong fluorescence was observed on the membrane of immature oocytes and apical surface of 2-cell embryos. In 8-cell embryos and morula, it could be observed throughout the cytoplasm. It was largely distributed in trophectoderm but little in inner cell mass at blastocyst stage. In IGF-IR experiments immunofluorescence was detected on the whole cell membranes of immature and mature oocytes, 2-cell and 8-cell embryos. Immunofluorescence of morula was observed in the whole cytoplasm of blastomere. In blastocysts, it was most distributed in trophectoderm but few in inner cell mass. It was proved that the expressions of the IGF-Ⅰ and IGF- IR were consecutive during preimplantation embryo development. Their expression throughout the whole cytoplasm in morula suggests that it could play an important role in the next differentiation and development.